Cephem compounds and processes for preparation thereof

ABSTRACT

The present invention relates to new cephem compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising the same and anti-bacterial methods of using the same. The subject cephem compounds comprise a thiadiazine ring bonded to the cephem through an iminoacetamido chain and a pyarzolium ring bonded to the cephem through a methylene chain.

This application is a Continuation of application Ser. No. 08/459,631,filed on Jun. 2, 1995, now abandoned, which is a Continuation ofapplication Ser. No. 08/135,583, filed on Oct. 13, 1993, now abandoned,which is a Continuation of application Ser. No. 07/993,615, filed onDec. 21, 1992, now abandoned, which is a Continuation-in-Part ofapplication Ser. No. 07/683,473, filed on Apr. 9, 1991, now U.S. Pat.No. 5,210,080, issued on May 11, 1993, which is a Continuation ofapplication Ser. No. 07/238,136, filed on Aug. 30, 1988, now abandoned.

The present invention relates to new cephem compounds andpharmaceutically acceptable salts thereof. More particularly, it relatesto new cephem compounds and pharmaceutically acceptable salts thereof,which have antimicrobial activities, to processes for preparationthereof, to pharmaceutical composition comprising the same, and to amethod for treating infectious diseases in human being and animals.

Accordingly, one object of the present invention is to provide thecephem compounds and pharmaceutically acceptable salts thereof, whichare highly active against a number of pathogenic microorganisms.

Another object of the present invention is to provide processes for thepreparation of the cephem compounds and salts thereof.

A further object of the present invention is to provide pharmaceuticalcomposition comprising, as an active ingredient, said cephem compoundsor their pharmaceutically acceptable salts.

Still further object of the present invention is to provide a method fortreating infectious diseases caused by pathogenic microorganisms, whichcomprises administering said cephem compounds to infected human being oranimals.

The object cephem compounds of the present invention are novel and canbe represented by the following general formula [I]: ##STR1## wherein R¹is amino or protected amino,

R² is lower alkyl, cyclo(lower)alkyl or lower alkenyl,

R³ is lower alkyl, hydroxy(lower)alkyl or protected hydroxy(lower)alkyl,

R⁴ is amino, protected amino, lower alkylamino, protected loweralkylamino, carboxy(lower)alkylamino, N-[protectedcarboxy(lower)alkyl]amino, and

R⁷ is hydrogen or lower alkyl.

As to the object compound [I], the following points are to be noted.

That is, the object compound [I] includes syn isomer, anti isomer and amixture thereof. Syn isomer means one geometrical isomer having thepartial structure represented by the following formula: ##STR2##(wherein R¹ and R² are each as defined above) and anti isomer means theother geometrical isomer having the partial structure represented by thefollowing formula: ##STR3## (wherein R¹ and R² are each as definedabove), and all of such geometrical isomers and mixture thereof areincluded within the scope of this invention.

In the present specification and claim, the partial structure of thesegeometrical isomers and mixture thereof are represented for convenientsake by the following formula: ##STR4## (wherein R¹ and R² are each asdefined above).

Another point to be noted is that the pyrazolio moiety of the compound[I] can also exist in the tautomeric form, and such tautomericequilibrium can be represented by the following schemes. ##STR5##(wherein R³, R⁴ and R⁷ are each as defined above)

Both of the above tautomeric isomers are included within the scope ofthe present invention, and in the present specification and claim,however, the object compound [I] is represented for the convenient sakeby one expression of the pyrazolio group of the formula (A).

The cephem compound [I] of the present invention can be prepared byprocesses as illustrated in the following. ##STR6## wherein R¹, R², R³,R⁴ and R⁷ are each as defined above,

R_(c) ⁴ is N-[protected carboxy(lower)alkyl]amino,

R_(d) ⁴ is carboxy(lower)alkylamino,

R_(a) ³ is protected hydroxy(lower)alkyl, and

R_(b) ³ is hydroxy(lower)alkyl.

The starting compound [II] can be prepared by the following processes.##STR7## wherein R³, R⁴ and R⁷ are each as defined above,

R⁵ is protected amino,

R⁶ is protected carboxy,

Y is a leaving group,

X.sup.⊖ is an anion,

R_(a) ⁴ is protected amino or protected lower alkylamino, and

R_(b) ⁴ is amino or lower alkylamino.

The starting compound [V] or a salt thereof can be prepared by themethods disclosed in the Preparations 1 to 8 described later or similarmanners thereto.

In the above and subsequent descriptions of this specification, suitableexamples of the various definitions are explained in detail as follows:

The term "lower" is intended to mean 1 to 6 carbon atom(s), unlessotherwise indicated.

Suitable "amino protective group" in the "protected amino" and"protected lower alkylamino" may be an acyl group as mentioned below,substituted or unsubstituted ar(lower)alkylidene [e.g. benzylidene,hydroxybenzylidene, etc.], ar(lower)alkyl such as mono or di ortriphenyl(lower)alkyl [e.g. benzyl, phenethyl, benzhydryl, trityl,etc.], or the like.

Suitable "acyl" may be lower alkanoyl [e.g. formyl, acetyl, propionyl,hexanoyl, pivaloyl, etc.], mono (or di or tri)halo(lower)alkanoyl [e.g.chloroacetyl, trifluoroacetyl, etc.], lower alkoxycarbonyl [e.g. methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, tert-pentyloxycarbonyl,hexyloxycarbonyl, etc.], carbamoyl, aroyl [e.g. benzoyl, toluoyl,naphthoyl, etc.], ar(lower)alkanoyl [e.g. phenylacetyl, phenylpropionyl,etc.], aryloxycarbonyl [e.g. phenoxycarbonyl, naphthyloxycarbonyl,etc.], aryloxy(lower)alkanoyl [e.g. phenoxyacetyl, phenoxypropionyl,etc.], arylglyoxyloyl [e.g. phenylglyoxyloyl, naphthylglyoxyloyl, etc.],ar(lower)alkoxycarbonyl which may have suitable substituent(s) [e.g.benzyloxycarbonyl, phenethyloxycarbonyl, p-nitrobenzyloxycarbonyl,etc.], or the like.

Suitable "lower alkyl" and "lower alkyl moieties" in the"hydroxy(lower)alkyl", "protected hydroxy(lower)alkyl", "loweralkylamino", "protected lower alkylamino", "carboxy(lower)alkylamino"and "N-[protected carboxy(lower)alkyl]amino", may be a straight orbranched one such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl,tert-butyl, pentyl, hexyl or the like.

Suitable "cyclo(lower)alkyl" may be one having 3 to 6 carbon atoms suchas cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

Suitable "lower alkenyl" may be a straight or branched one such asvinyl, allyl, 1-propenyl, isopropenyl, butenyl, isobutenyl, pentenyl,hexenyl, methylpropenyl or the like.

Suitable "protected hydroxy" in the "protected hydroxy(lower)alkyl" maybe acyloxy group or the like. Suitable "acyl moiety" in the "acyloxy"may be lower alkanoyl [e.g. formyl, acetyl, propionyl, hexanoyl,pivaloyl, etc.], mono (or di or tri)halo(lower)alkanoyl [e.g.chloroacetyl, trifluoroacetyl, etc.], lower alkoxycarbonyl [e.g.methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl,tert-pentyloxycarbonyl, hexyloxycarbonyl, etc.], carbamoyl or the like.

Suitable "protected carboxy" and "protected carboxy moiety" in the term"N-[protected carboxy(lower)alkyl]amino" may be an esterified carboxygroup, or the like, and concrete examples of the ester moiety in saidesterified carboxy group may be the ones such as lower alkyl ester [e.g.methyl ester, ethyl ester, propyl ester, isopropyl ester, butyl ester,isobutyl ester, tert-butyl ester, pentyl ester, hexyl ester,1-cyclopropylethyl ester, etc.] which may have suitable substituent(s),for example, lower alkanoyloxy(lower)alkyl ester [e.g. acetoxymethylester, propionyloxymethyl ester, butyryloxymethyl ester,valeryloxymethyl ester, pivaloyloxymethyl ester, 1-acetoxyethyl ester,1-propionyloxyethyl ester, pivaloyloxymethyl ester, 2-propionyloxyethylester, hexanoyloxymethyl ester, etc.], lower alkanesulfonyl(lower)alkylester [e.g. 2-mesylethyl ester, etc.] or mono(or di ortri)halo(lower)alkyl ester [e.g. 2-iodoethyl ester, 2,2,2-trichloroethylester, etc.]; lower alkenyl ester [e.g. vinyl ester, allyl ester, etc.];lower alkynyl ester [e.g. ethynyl ester, propynyl ester, etc.];ar(lower)alkyl ester which may have suitable substituent(s) [e.g. benzylester, 4-methoxybenzyl ester, 4-nitrobenzyl ester, phenethyl ester,trityl ester, benzhydryl ester, bis(methoxyphenyl)methyl ester,3,4-dimethoxybenzyl ester, 4-hydroxy-3,5-di-tert-butylbenzyl ester,etc.]; aryl ester which may have suitable substituent(s) [e.g. phenylester, 4-chlorophenyl ester, tolyl ester, 4-tert-butylphenyl ester,xylyl ester, mesityl ester, cumenyl ester, etc.]; or the like.

Suitable "leaving group" may be halogen [e.g. chlorine, bromine, iodine,etc.], acyloxy such as sulfonyloxy [e.g. benzenesulfonyloxy, tosyloxy,mesyloxy, etc.], lower alkanoyloxy [e.g. acetyloxy, propionyloxy, etc.],or the like.

Suitable "anion" may be formate, acetate, trifluoroacetate, maleate,tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate,chloride, bromide, iodide, sulfate, phosphate, or the like.

Suitable pharmaceutically acceptable salts of the object compound [I]are conventional non-toxic salts and include a metal salt such as analkali metal salt [e.g. sodium salt, potassium salt, etc.] and analkaline earth metal salt [e.g. calcium salt, magnesium salt, etc.], anammonium salt, an organic base salt [e.g. trimethylamine salt,triethylamine salt, pyridine salt, picoline salt, dicyclohexylaminesalt, N,N'-dibenzylethylenediamine salt, etc.], an acid addition saltsuch as an organic acid addition salt [e.g. formate, acetate,trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate,toluenesulfonate, etc.], an inorganic acid addition salt [e.g.hydrochloride, hydrobromide, sulfate, phosphate, etc.], a salt with anamino acid [e.g. arginine salt, aspartic acid salt, glutamic acid salt,etc.], and the like.

Preferred embodiments of the object compound [I] are as follows.

Preferred embodiment of R¹ is amino,

R² is lower alkyl [more preferably (C₁ -C₄)alkyl, most preferablyethyl], cyclo(lower)alkyl [more preferably cyclo(C₄ -C₆)alkyl] or loweralkenyl [more preferably (C₂ -C₆)alkenyl, most preferably propenyl],

R³ is lower alkyl [more preferably (C₁ -C₄)alkyl, most preferablymethyl], hydroxy(lower)alkyl [more preferably hydroxy (C₁ -C₄ )alkyl,most preferably hydroxyethyl] or acyloxy(lower)alkyl [more preferablycarbamoyloxy(lower)alkyl or lower alkanoyloxy(lower)alkyl, mostpreferably carbamoyloxy (C₁ -C₄)alkyl or lower alkanoyloxy (C₁-C₄)alkyl],

R⁴ is amino, acylamino [more preferably carbamoylamino or loweralkanoylamino], lower alkylamino, carboxy(lower)alkylamino orN-[protected carboxy(lower)alkyl]amino [more preferably esterifiedcarboxy(lower)alkylamino, most preferably loweralkoxycarbonyl(lower)alkylamino],

R⁷ is hydrogen or lower alkyl [more preferably (C₁ -C₄)alkyl, mostpreferably methyl].

The processes for preparing the object compound of the present inventionare explained in detail in the following.

Process 1

The compound [I] or a salt thereof can be prepared by reacting thecompound [II] or its reactive derivative at the amino group or a saltthereof with the compound [III] or its reactive derivative at thecarboxy group or a salt thereof.

Suitable reactive derivative at the amino group of the compound [II] mayinclude Schiff's base type imino or its tautomeric enamine type isomerformed by the reaction of the compound [II] with a carbonyl compoundsuch as aldehyde, ketone or the like; a silyl derivative formed by thereaction of the compound [II] with a silyl compound such asbis(trimethylsilyl)acetamide, mono(trimethylsilyl)acetamide [e.g.N-(trimethylsilyl)acetamide], bis(trimethylsilyl)urea or the like; aderivative formed by reaction of the compound [II] with phosphorustrichloride or phosgene, and the like.

Suitable salts of the compound [II] and its reactive derivative can bereferred to the ones as exemplified for the compound [I].

Suitable reactive derivative at the carboxy group of the compound [III]may include an acid halide, an acid anhydride, an activated amide, anactivated ester, and the like. Suitable examples of the reactivederivatives may be an acid chloride; an acid azide; a mixed acidanhydride with an acid such as substituted phosphoric acid [e.g.dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid,dibenzylphosphoric acid, halogenated phosphoric acid, etc.],dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, sulfuricacid, sulfonic acid [e.g. methanesulfonic acid, etc.], aliphaticcarboxylic acid [e.g. acetic acid, propionic acid, butyric acid,isobutyric acid, pivalic acid, pentanoic acid, isopentanoic acid,2-ethylbutyric acid, trichloroacetic acid, etc.] or aromatic carboxylicacid [e.g. benzoic acid, etc.]; a symmetrical acid anhydride; anactivated amide with imidazole, 4-substituted imidazole,dimethylpyrazole, triazole or tetrazole; or an activated ester [e.g.cyanomethyl ester, methoxymethyl ester, dimethyliminomethyl [(CH₃)₂ ⁺N═CH--] ester, vinyl ester, propargyl ester, p-nitrophenyl ester,2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester,mesylphenyl ester, phenylazophenyl ester, phenyl thioester,p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester,pyranyl ester, pyridyl ester, piperidyl ester, 8-quinolyl thioester,etc.], or an ester with a N-hydroxy compound [e.g.N,N-dimethylhydroxylamine, 1-hydroxy-2-(1H)-pyridone,N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxy-1H-benzotriazole,etc.], and the like. These reactive derivatives can optionally beselected from them according to the kind of the compound [III] to beused.

Suitable salts of the compound [III] and its reactive derivative can bereferred to the ones as exemplified for the compound [I].

The reaction is usually carried out in a conventional solvent such aswater, alcohol [e.g. methanol, ethanol, etc.], acetone, dioxane,acetonitrile, chloroform, methylene chloride, ethylene chloride,tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine or anyother organic solvent which does not adversely influence the reaction.These conventional solvent may also be used in a mixture with water.

In this reaction, when the compound [III] is used in a free acid form orits salt form, the reaction is preferably carried out in the presence ofa conventional condensing agent such as N,N'-dicyclohexylcarbodiimide;N-cyclohexyl-N'-morpholinoethylcarbodiimide;N-cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide;N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide;N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide;N,N'-carbonylbis-(2-methylimidazole);pentamethyleneketene-N-cyclohexylimine;diphenylketene-N-cyclohexylimine; ethoxyacetylene;1-alkoxy-1-chloroethylene; trialkyl phosphite; ethyl polyphosphate;isopropyl polyphosphate; phosphorus oxychloride (phosphoryl chloride);phosphorus pentachloride; phosphorus trichloride; thionyl chloride;oxalyl chloride; lower alkyl haloformate [e.g. ethyl chloroformate,isopropyl chloroformate, etc.]; triphenylphosphine;2-ethyl-7-hydroxybenzisoxazolium salt;2-ethyl-5-(m-sulfophenyl)isoxazolium hydroxide intramolecular salt;1-(p-chlorobenzenesulfonyloxy)-6-chloro-1H-benzotriazole; so-calledVilsmeier reagent prepared by the reaction of N,N-dimethylformamide withthionyl chloride, phosgene, trichloromethyl chloroformate, phosphorusoxychloride, etc.; or the like.

The reaction may also be carried out in the presence of an inorganic ororganic base such as an alkali metal bicarbonate, tri(lower)alkylamine,pyridine, N-(lower)alkylmorpholine, N,N-di(lower)alkylbenzylamine, orthe like.

The reaction temperature is not critical, and the reaction is usuallycarried out under cooling to warming.

Process 2

The compound [Ib] or a salt thereof can be prepared by subjecting thecompound [Ia] or a salt thereof to elimination reaction of the carboxyprotective group.

Suitable method of this elimination reaction may include conventionalone such as hydrolysis, reduction and the like.

(i) For Hydrolysis:

The hydrolysis is preferably carried out in the presence of a base or anacid including Lewis acid. Suitable base may include an inorganic baseand an organic base such as an alkali metal [e.g. sodium, potassium,etc.], an alkaline earth metal [e.g. magnesium, calcium, etc.], thehydroxide or carbonate or bicarbonate thereof, trialkylamine [e.g.trimethylamine, triethylamine, etc.], picoline,1,5-diazabicyclo[4.3.0]non-5-ene, 1,4-diazabicyclo[2.2.2]octane,1,8-diazabicyclo[5.4.0]undec-7-ene, or the like.

Suitable acid may include an organic acid [e.g. formic acid, aceticacid, propionic acid, trichloroacetic acid, trifluoroacetic acid, etc.]and an inorganic acid [e.g. hydrochloric acid, hydrobromic acid,sulfuric acid, hydrogen chloride, hydrogen bromide, etc.]. Theelimination using Lewis acid such as trihaloacetic acid [e.g.trichloroacetic acid, trifluoroacetic acid, etc.] or the like ispreferably carried out in the presence of cation trapping agents [e.g.anisole, phenol, etc.].

The reaction is usually carried out in a solvent such as water, analcohol [e.g. methanol, ethanol, etc.], methylene chloride,tetrahydrofuran, a mixture thereof or any other solvent which does notadversely influence the reaction. A liquid base or acid can be also usedas the solvent. The reaction temperature is not critical and thereaction is usually carried out under cooling to warming.

(ii) For Reduction:

Reduction is carried out in a conventional manner, including chemicalreduction and catalytic reduction.

Suitable reducing agents to be used in chemical reduction are acombination of a metal (e.g. tin, zinc, iron, etc.) or metallic compound(e.g. chromium chloride, chromium acetate, etc.) and an organic orinorganic acid (e.g. formic acid, acetic acid, propionic acid,trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid,hydrobromic acid, etc.).

Suitable catalysts to be used in catalytic reduction are conventionalones such as platinum catalysts (e.g. platinum plate, spongy platinum,platinum black, colloidal platinum, platinum oxide, platinum wire,etc.), palladium catalysts (e.g. spongy palladium, palladium black,palladium oxide, palladium on carbon, colloidal palladium, palladium onbarium sulfate, palladium on barium carbonate, etc.), nickel catalysts(e.g. reduced nickel, nickel oxide, Raney nickel, etc.), cobaltcatalysts (e.g. reduced cobalt, Raney cobalt, etc.), iron catalysts(e.g. reduced iron, Raney iron, etc.), copper catalysts (e.g. reducedcopper Raney copper, Ullman copper, etc.) and the like. The reduction isusually carried out in a conventional solvent which does not adverselyinfluence the reaction such as water, methanol, ethanol, propanol,N,N-dimethylformamide, or a mixture thereof. Additionally, in case thatthe above-mentioned acids to be used in chemical reduction are inliquid, they can also be used as a solvent. Further, a suitable solventto be used in catalytic reduction may be the above-mentioned solvent,and other conventional solvent such as diethyl ether, dioxane,tetrahydrofuran, etc., or a mixture thereof.

The reaction temperature of this reduction is not critical and thereaction is usually carried out under cooling to warming.

Process 3

The compound [Id] or a salt thereof can be prepared by subjecting thecompound [Ic] or a salt thereof to elimination reaction of the hydroxyprotective group. This reaction can be carried out in a similar mannerto that of the aforementioned Process 2, and therefore the reagents tobe used and the reaction conditions (e.g., solvent, reactiontemperature, etc.) can be referred to those of the Process 2.

Processes A and B for the preparation of the starting compounds areexplained in detail in the following.

Process A-1

The compound [VI] or a salt thereof can be prepared by reacting thecompound [IV] or a salt thereof with the compound [V] or a salt thereof.

Suitable salts of the compounds [V] and [VI] can be referred to the onesas exemplified for the compound [I].

The present reaction may be carried out in a solvent such as water,phosphate buffer, acetone, chloroform, acetonitrile, nitrobenzene,methylene chloride, ethylene chloride, formamide, N,N-dimethylformamide,methanol, ethanol, diethyl ether, tetrahydrofuran, dimethyl sulfoxide,or any other organic solvent which does not adversely affect thereaction, preferably in ones having strong polarities. Among thesolvents, hydrophilic solvents may be used in a mixture with water. Whenthe compound [V] is in liquid, it can also be used as a solvent. Thereaction is preferably conducted in the presence of a base, for example,inorganic base such as alkali metal hydroxide, alkali metal carbonate,alkali metal bicarbonate, organic base such as trialkylamine, and thelike. The reaction temperature is not critical, and the reaction isusually carried out at ambient temperature, under warming or underheating. The present reaction is preferably carried out in the presenceof alkali metal halide [e.g. sodium iodide, potassium iodide, etc.],alkali metal thiocyanate [e.g. sodium thiocyanate, potassiumthiocyanate, etc.] or the like.

Anion X.sup.⊖ may be the one derived from a leaving group Y and may bethe other one converted therefrom by a conventional method.

Process A-2

The compound [II] or a salt thereof can be prepared by subjecting thecompound [VI] or a salt thereof to elimination reaction of the aminoprotective group in R⁵ and the carboxy protective group in R⁶.

This reaction is carried out in accordance with a conventional methodsuch as hydrolysis or the like.

The hydrolysis is preferably carried out in the presence of a base or anacid including Lewis acid. Suitable base may include an inorganic baseand an organic base such as an alkali metal [e.g. sodium, potassium,etc.], an alkaline earth metal [e.g. magnesium, calcium, etc.], thehydroxide or carbonate or bicarbonate thereof, trialkylamine [e.g.trimethylamine, triethylamine, etc.], picoline,1,5-diazabicyclo-[4.3.0]non-5-ene, 1,4-diazabicyclo[2.2.2]octane,1,8-diazabicyclo[5.4.0]undec-7-ene, or the like. Suitable acid mayinclude an organic acid [e.g. formic acid, acetic acid, propionic acid,trichloroacetic acid, trifluoroacetic acid, etc.] and an inorganic acid[e.g. hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogenchloride, hydrogen bromide, etc.]. The elimination using Lewis acid suchas trihaloacetic acid [e.g. trichloroacetic acid, trifluoroacetic acid,etc.] or the like is preferably carried out in the presence, of cationtrapping agents [e.g. anisole, phenol, etc.].

The reaction is usually carried out in a solvent such as water, analcohol [e.g. methanol, ethanol, etc.], methylene chloride,tetrahydrofuran, a mixture thereof or any other solvent which does notadversely influence the reaction. A liquid base or acid can be also usedas the solvent. The reaction temperature is not critical and thereaction is usually carried out under cooling to warming.

The present invention includes within the scope of the invention thecase that protected amino in R⁴ is transformed into amino during thisreaction.

Process B

The compound [IIb] or a salt thereof can be prepared by subjecting thecompound [IIa] to elimination reaction of the amino protective group inR_(a) ⁴. This reaction can be carried out in a similar manner to that ofthe aforementioned Process A-2, and therefore the reagents to be usedand the reaction conditions (e.g., solvent, reaction temperature, etc.)can be referred to those of the Process A-2. The present inventionincludes within the scope of the invention the case that protectedhydroxy(lower)alkyl in R³ is transformed into hydroxy(lower)alkyl duringthis reaction.

The object compound [I] and pharmaceutically acceptable salts thereofare novel and exhibit high antimicrobial activity, inhibiting the growthof a wide variety of pathogenic microorganisms including Gram-positiveand Gram-negative microorganisms and are useful as antimicrobial agents.

Now in order to show the utility of the object compound [I], the testdata on MIC (minimal inhibitory concentration) of a representativecompound of this invention are shown in the following.

Test method:

In vitro antibacterial activity was determined by the two-foldagar-plate dilution method as described below.

One loopful of an overnight culture of each test strain inTrypticase-soy broth (10⁶ viable cells per ml) was streaked on heartinfusion agar (HI-agar) containing graded concentrations ofrepresentative test compound, and the minimal inhibitory concentration(MIC) was expressed in terms of μg/ml after incubation at 37° C. for 20hours.

Test compounds:

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer) (hereinafter referred to as Compound A)

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer) (hereinafter referred to as Compound B)

Test results:

    ______________________________________                                        Test results:                                                                                 MIC (μg/ml)                                                Test            Test compound                                                 strain          A      B                                                      ______________________________________                                        E. coli 31      0.05   ≦0.025                                          ______________________________________                                    

For therapeutic administration, the object compound [I] andpharmaceutically acceptable salts thereof of the present invention areused in the form of conventional pharmaceutical preparation whichcontains said compound as an active ingredient, in admixture withpharmaceutically acceptable carriers such as an organic or inorganicsolid or liquid excipient which is suitable for oral, parenteral andexternal (topical) administration. The pharmaceutical preparations maybe in solid form such as tablet, granule, powder, capsule, or liquidform such as solution, suspension, syrup, emulsion, lemonade and thelike.

If needed, there may be included in the above preparations auxiliarysubstances, stabilizing agents, wetting agents and other commonly usedadditives such as lactose, citric acid, tartaric acid, stearic acid,magnesium stearate, terra alba, sucrose, corn starch, talc, gelatin,agar, pectin, peanut oil, olive oil, cacao butter, ethylene glycol, andthe like.

While the dosage of the compound [I] may vary from and also depend uponthe age, conditions of the patient, a kind of diseases, a kind of thecompound [I] to be applied, etc. In general amounts between 1 mg andabout 4,000 mg or even more per day may be administered to a patient. Anaverage single dose of about 50 mg, 100 mg, 250 mg, 500 mg, 1000 mg,2000 mg of the object compounds [I] of the present invention may be usedin treating diseases infected by pathogenic microorganisms.

The following Preparations and Examples are given for the purpose ofillustrating the present invention in more detail.

Preparation 1

(1) A mixture of acetic anhydride (11.13 ml) and formic acid (5.93 ml)was stirred at ambient temperature for 30 minutes. To this solution wasadded 5-amino-1-(2-hydroxyethyl)pyrazole (5 g) under ice-cooling, andthe mixture was stirred at 30°-40° C. for 1 hour. The reaction mixturewas poured into a mixture of water, tetrahydrofuran and ethyl acetateand adjusted to pH 6 with aqueous sodium bicarbonate. The organic layerwas separated, and the aqueous layer was extracted with a mixture oftetrahydrofuran and ethyl acetate for three times. The organic layerswere combined, dried over magnesium sulfate and evaporated in vacuo togive 5-formamido-1-(2-formyloxyethyl)pyrazole (5.18 g).

IR (Nujol): 3180, 1705, 1660 cm⁻¹

NMR (DMSO-d₆, δ): 4.21-4.61 (4H, m), 6.11 and 6.34 (1H, each d, J=3 Hz),7.47 (1H, d, J=3 Hz), 8.00 (1H, s), 8.33 (1H, s).

The following compound was obtained according to a similar manner tothat of Preparation 1(1).

(2) 5-Formamido-1-(2-formyloxyethyl)-4-methylpyrazole

IR (Nujol): 3180, 1715, 1660 cm⁻¹

NMR (DMSO-d₆, δ): 1.81 and 1.86 (3H, each s), 4.01-4.48 (4H, m), 7.25and 7.40 (1H, each s), 8.06 (1H, s), 8.22 and 9.13 (1H, each s).

Preparation 2

5-Amino-1-(2-hydroxyethyl)pyrazole (10 g) was added to a mixture ofacetic acid (50 ml) and water (100 ml). A solution of potassium cyanate(25.5 g) in water (80 ml) was added dropwise thereto under stirring at34° C. The mixture was stirred at room temperature overnight.

The reaction mixture was added to ethyl acetate (200 ml). The mixturewas adjusted to pH 8.0 with potassium carbonate and extracted withtetrahydrofuran. The extract was dried over magnesium sulfate, and theorganic solvent was evaporated in vacuo to give an oily product. Acetonewas added to the said oily product to give5-carbamoylamino-1-(2-hydroxyethyl)pyrazole (3.76 g) as amorphous solid.

IR (Nujol): 3400, 3200, 1670, 1570 cm⁻¹

NMR (DMSO-d₆, δ): 3.64 (2H, t, J=6 Hz), 3.97 (2H, t, J=6 Hz), 6.02 and6.13 (1H, each d, J=2 Hz), 7.20 and 7.27 (1H, each d, J=2 Hz).

Preparation 3

Formic acid (2.09 ml) was added to acetic anhydride (4.17 ml) at roomtemperature and the mixture was stirred for 30 minutes at the sametemperature. 5-Carbamoylamino-1-(2-hydroxyethyl)pyrazole (3.76 g) wasadded thereto under stirring and ice-cooling. The mixture was stirredfor 2 hours at room temperature. The reaction mixture was evaporated invacuo and diisopropyl ether was added thereto to give5-carbamoylamino-1-(2-formyloxyethyl)pyrazole (4.15 g).

mp: 102°-104° C.

IR (Nujol): 3400, 3200, 1710, 1660, 1560, 1170 cm⁻¹

NMR (D₂ O, δ): 4.20 (2H, t, J=5 Hz), 4.36 (2H, t, J=5 Hz), 5.96 (1H, s),6.02 and 6.13 (1H, each d, J=2 Hz), 7.24 and 7.32 (1H, each d, J=2 Hz)8.06 and 8.24 (1H, each s).

Preparation 4

1-(2-Hydroxyethyl)-5-aminopyrazole (5 g) was added to acetic anhydride(14.7 ml) under stirring and ice-cooling, and pyridine (6.3 ml) wasadded thereto. The mixture was stirred for 2 hours at 25° C. Thereaction mixture was added to a mixture of ethyl acetate (50 ml) andbrine (50 ml). Then, the mixture was adjusted to pH 7.0 with an aqueoussolution of sodium bicarbonate. The aqueous layer was extracted with amixture of ethyl acetate and tetrahydrofuran. The extract was dried overmagnesium sulfate. The magnesium sulfate was filtered off, and thefiltrate was evaporated under reduced pressure to give1-(2-acetoxyethyl)-5-acetylaminopyrazole (5.98 g).

mp: 83°-84° C.

IR (Nujol): 3270, 1750, 1670, 1565 cm⁻¹

NMR (DMSO-d₆, δ): 1.93 (3H, s), 2.03 (3H, s), 4.22 (4H, br s), 6.13 (1H,d, J=2 Hz), 7.32 (1H, d, J=2 Hz), 9.76 (1H, s).

Preparation 5

A mixture of acetic anhydride (44.5 ml) and formic acid (22.3 ml) wasstirred at ambient temperature for an hour. To this mixture was added1-(2-hydroxyethyl)-5-aminopyrazole (30 g) at 0°-10° C., and the mixturewas stirred under ice-cooling for 30 minutes. The mixture was pouredinto ice-cold water, adjusted to pH 10.5 with 40% aqueous potassiumcarbonate, and stirred under ice-cooling for 30 minutes. The mixture wasextracted with a mixture of tetrahydrofuran and ethyl acetate 6 times.The organic layer was dried over magnesium sulfate and evaporated invacuo to give 1-(2-hydroxyethyl)-5-formamidopyrazole (30.8 g).

IR (Nujol): 3230, 1695, 1570, 1540 cm⁻¹

NMR (DMSO-d₆, δ): 3.62-3.95 (2H, m), 3.98-4.32 (2H, m), 6.22 and 6.36(1H, each d, J=3 Hz), 7.42 (1H, d, J=3 Hz), 8.32 and 8.36 (1H, each s).

Preparation 6

To a suspension of 5-formamido-1-(2-hydroxyethyl)pyrazole (1 g) inacetonitrile (50 ml) was added dropwise chlorosulfonyl isocyanate (0.77ml) at -15° C.˜-20° C. The mixture was stirred for 3 hours underice-cooling. To the reaction mixture was added water (1 ml) and kept tostand overnight. The solution was adjusted to pH 7.5 with 5N-sodiumhydroxide solution and then adjusted to pH 8.5 with 1N-sodium hydroxidesolution. The organic layer was separated and the aqueous layer wasextracted with tetrahydrofuran. The extract and said organic layer werecombined and dried over magnesium sulfate. The solvent was distilled offand the residue was crystallized from ethyl acetate to give5-amino-1-(2-carbamoyloxyethyl)pyrazole (0.60 g).

NMR (DMSO-d₆, δ): 3.83-4.35 (4H, m), 4.80-5.18 (2H, broad s), 5.32 (1H,d, J=3 Hz), 6.33-6.87 (2H, broad s), 7.08 (1H, d, J=3 Hz).

Preparation 7

5-Formamido-1-(2-carbamoyloxyethyl) pyrazole (3.69 g) was obtained from5-amino-1-(2-carbamoyloxyethyl)pyrazole (3.3 g) according to a similarmanner to that of Preparation 5.

NMR (DMSO-d₆, δ): 4.22 (4H, s), 6.17-6.40 (1H, m), 6.40-6.63 (2H, m),7.30-7.53 (1H, m), 8.13-8.47 (1H, m).

Preparation 8

(1) To a solution of 5-formamido-1-methylpyrazole inN,N-dimethylformamide (50 ml) was sodium hydride (1.6 g) underice-cooling. Then, to the mixture was added methyl iodide (2.5 ml) atthe same condition. The mixture was stirred for 1 hour underice-cooling. To the reaction mixture was added a mixture of ethylacetate (500 ml) and water (100 ml). The organic layer was separated,and the aqueous layer was extracted with ethyl acetate (50 ml×2). Theorganic solution was dried over magnesium sulfate. The solvent wasdistilled off and the residue was subjected to a column chromatographyon silica gel using a mixture of ethyl acetate and diisopropyl ether(3:1) as an eluent. Fractions containing the object compound werecollected and evaporated in vacuo to give5-(N-formyl-N-methylamino)-1-methylpyrazole (2.5 g).

IR (Nujol): 1660-1680, 1550, 1320 cm⁻¹

NMR (DMSO-d₆, δ): 3.07 (3H, s), 3.67 (3H, s), 6.28 (1H, d, J=2 Hz), 7.44(1H, d, J=2 Hz), 8.20 (1H, s).

The following compound was obtained according to a similar manner tothat of Preparation 8(1).

(2) 5-(N-Formyl-N-methoxycarbonylmethylamino)-1-methylpyrazole

NMR (CDCl₃, δ): 3.68 (3H, s), 3.77 (3H, s), 4.26 (2H, s), 6.16 (1H, d,J=3 Hz), 7.39 (1H, d, J=3 Hz), 8.15 (1H, s).

Preparation 9

(1) To a mixture of benzhydryl7β-tert-butoxycarbonylamino-3-chloromethyl-3-cephem-4-carboxylate (20 g)and sodium iodide (5.82 g) in N,N-dimethylformamide (20 ml) was added5-formamido-1-(2-formyloxyethyl) pyrazole (21.34 g) at ambienttemperature. After being stirred for 24 hours at the same temperature,the mixture was poured into a mixture of water and ethyl acetate. Theorganic layer was separated and washed with water, aqueous sodiumchloride solution, and dried over magnesium sulfate. The solution wasevaporated in vacuo to give benzhydryl7β-tert-butoxycarbonylamino-3-[3-formamido-2-(2-formyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylateiodide (29.6 g).

IR (Nujol): 1780, 1720 cm⁻¹

NMR (DMSO-d₆, δ): 1.49 (9H, s), 3.43 (2H, br s), 4.14-4.38 (2H, m),4.52-4.73 (2H, m), 5.15 (1H, d, J=5 Hz), 5.40 (2H, br s), 5.67 (1H, dd,J=5, 8 Hz), 6.88 (1H, s), 7.02 (1H, d, J=3 Hz), 7.18-7.52 (10H, m), 7.94(1H, d, J=8 Hz), 7.99 (1H, s), 8.27 (1H, d, J=3 Hz), 8.51 (1H, br s).

The following compounds were obtained according to a similar manner tothat of Preparation 9 (1).

(2) Benzhydryl7β-tert-butoxycarbonylamino-3-[3-formamido-2-(2-formyloxyethyl)-4-methyl-1-pyrazolio]-methyl-3-cephem-4-carboxylateiodide

IR (Nujol): 3250, 1780, 1710, 1680 cm⁻¹

NMR (DMSO-d₆, δ): 1.53 (9H, s), 1.97 (3H, s), 3.51 (2H, br s), 4.04-4.42(2H, m), 4.52-4.78 (2H, m), 5.08 (1H, d, J=5 Hz), 5.39 (2H, br s), 5.61(1H, dd, J=5, 8 Hz), 6.86 (1H, s), 7.08-7.52 (10H, m), 7.93 (1H, s),8.18 (1H, s), 8.34 (1H, s), 9.12 (1H, s).

(3) Benzhydryl7β-tert-butoxycarbonylamino-3-[3-carbamoylamino-2-(2-formyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylateiodide

IR (Nujol): 3300, 1780, 1710 cm⁻¹

NMR (DMSO-d₆, δ): 1.43 (9H, s), 3.5-3.8 (2H, m), 4.1-4.6 (4H, m), 5.14(1H, d, J=5 Hz), 5.33 (2H, s), 5.60 (1H, dd, J=8 Hz, 5 Hz), 6.66 (1H,s), 6.86 (1H, d, J=3 Hz), 7.1-7.5 (10H, m), 7.93 (1H, d, J=8 Hz), 8.00(1H, s), 8.08 (1H, d, J=3 Hz).

(4) Benzhydryl7β-tert-butoxycarbonylamino-3-[3-acetamido-2-(2-acetoxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylateiodide

IR (Nujol): 1780, 1720, 1230 cm⁻¹

NMR (DMSO-d₆, δ): 1.41 (9H, s), 1.86 (3H, s), 2.25 (3H, s), 3.40 (2H, brs), 4-4.4 (4H, m), 5.12 (1H, d, J=5 Hz), 5.37 (2H, s), 5.60 (1H, dd, J=8Hz, 5 Hz), 6.85 (1H, s), 7.24 (1H, d, J=3 Hz), 7.1-7.6 (10H, m), 7.90(1H, d, J=8 Hz), 8.21 (1H, d, J=3 Hz), 11.17 (1H, s).

(5) Benzhydryl7β-tert-butoxycarbonylamino-3-[2-(2-carbamoyloxyethyl)-3-formamido-1-pyrazolio)methyl-3-cephem-4-carboxylateiodide

(6) Benzhydryl7-tert-butoxycarbonylamino-3-[3-(N-formyl-N-methylamino)-2-methyl-1-pyrazolio]methyl-3-cephem-4-carboxylateiodide

NMR (DMSO-d₆, δ): 1.38 (9H, s), 3.29 (3H, s), 3.68 (3H, s), 3.07-3.77(2H, m), 5.18 (1H, d, J=5 Hz), 5.35-5.75 (3H, m), 6.90 (1H, s), 7.01(1H, d, J=2 Hz), 7.08-7.60 (10H, m), 8.02 (1H, d, J=8 Hz), 8.35 (1H, s),8.43 (1H, d, J=2 Hz).

(7) Benzhydryl7-tert-butoxycarbonylamino-3-[3-(N-formyl-N-methoxycarbonylmethylamino)-2-methyl-1-pyrazolio]methyl-3-cephem-4-carboxylateiodide

IR (Nujol): 3300, 1780, 1620 cm⁻¹.

Preparation 10

(1) To a solution of benzhydryl7β-tert-butoxycarbonylamino-3-[3-formamido-2-(2-formyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylateiodide (29.5 g) and anisole (30 ml) in methylene chloride (90 ml) wasadded dropwise trifluoroacetic acid (60 ml) under ice-cooling. Afterbeing stirred for 1 hour at ambient temperature, the mixture was pouredinto a mixture of diisopropyl ether (600 ml) and ethyl acetate (600 ml).The resultant precipitate was collected by filtration to givedi(trifluoroacetic acid) salt of7β-amino-3-[3-formamido-2-(2-formyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(22.7 g).

IR (Nujol): 1780, 1715, 1660 cm⁻¹

NMR (DMSO-d₆, δ): 3.53 (2H, br s), 4.28-4.56 (2H, m), 4.78-4.99 (2H, m),5.29 (2H, br s), 5.53 (2H, br s), 7.14 (1H, d, J=3 Hz), 8.22 (1H, s),8.46 (1H, d, J=3 Hz), 8.63 (1H, s).

The following compounds were obtained according to a similar manner tothat of Preparation 10 (1).

(2) Di(trifluoroacetic acid) salt of7β-amino-3-[3-formamido-2-(2-formyloxyethyl)-4-methyl-1-pyrazolio]methyl-3-cephem-4-carboxylate

IR (Nujol): 1780, 1710, 1670 cm⁻¹

NMR (DMSO-d₆, δ): 1.98 (3H, s), 3.49 (2H, br s), 4.22-4.48 (2H, m),4.61-4.87 (2H, m), 5.18 (2H, br s), 5.46 (2H, br s), 8.05 (1H, s), 8.23(1H, s), 8.35 (1H, s).

(3) Di(trifluoroacetic acid) salt of7β-amino-3-[3-carbamoylamino-2-(2-formyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate

IR (Nujol): 3400, 1780, 1700, 1560 cm⁻¹

NMR (DMSO-d₆, δ): 3.46 (2H, br s), 4.2-4.6 (4H, m), 5.22 (2H, m), 5.40(2H, s), 6.92 (1H, d, J=3 Hz), 8.13 (1H, s), 8.20 (1H, d, J=3 Hz).

(4) Di(trifluoroacetic acid) salt of7β-amino-3-[3-acetamido-2-(2-acetoxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate

IR (Nujol): 1780, 1660, 1190 cm⁻¹

NMR (DMSO-d₆, δ): 1.95 (3H, s), 2.23 (3H, s), 3.46 (2H, br s), 4.1-4.4(4H, m), 5.20 (2H, m), 5.46 (2H, s), 7.01 (1H, d, J=3 Hz), 8.27 (1H, d,J=3 Hz), 11.17 (1H, s).

(5) Di(trifluoroacetic acid) salt of7β-amino-3-[2-(2-carbamoyloxyethyl)-3-formamido-1-pyrazolio]methyl-3-cephem-4-carboxylate

(6) Di(trifluoroacetic acid) salt of7β-amino-3-[3-(N-formyl-N-methylamino)-2-methyl-1-pyrazolio]methyl-3-cephem-4-carboxylate

NMR (D₂ O, δ): 3.46 (3H, s), 3.87, 4.00 (total 3H, each s), 3.13-3.77(2H, m), 5.05-5.47 (4H, m), 6.89 (1H, d, J=2 Hz), 8.32 (1H, d, J=2 Hz),8.36 (1H, s).

(7) Di(trifluoroacetic acid) salt of7β-amino-3-[3-(N-formyl-N-methoxycarbonylmethylamino)-2-methyl-1-pyrazolio]methyl-3-cephem-4-carboxylate

IR (Nujol): 3300, 1780, 1610 cm⁻¹.

Preparation 11

(1) Concentrated hydrochloric acid (5.67 ml) was added to a mixture ofdi(trifluoroacetic acid) salt of7β-amino-3-[3-formamido-2-(2-formyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(10 g) in methanol (50 ml) at ambient temperature. After being stirredat the same temperature for 3 hours, the mixture was added dropwise toethyl acetate (500 ml). The resultant precipitate was collected byfiltration to give7β-amino-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylatetrihydrochloride (6.1 g).

IR (Nujol): 3250, 1770, 1700, 1625 cm⁻¹

NMR (DMSO-d₆, δ): 3.43 (2H, br s), 3.52-3.88 (2H, m), 4.18-4.48 (2H, m),5.28 (2H, br s), 5.37 (2H, br s), 5.97 (1H, d, J=3 Hz), 8.18 (1H, d, J=3Hz).

The following compounds were obtained according to a similar manner tothat of Preparation 11(1).

(2)7β-Amino-3-[3-amino-2-(2-hydroxyethyl)-4-methyl-1-pyrazolio]methyl-3-cephem-4-carboxylatetrihydrochloride

NMR (DMSO-d₆, δ): 1.94 (3H, s), 3.39 (2H, br s), 3.47-3.78 (2H, m),4.06-4.42 (2H, m), 5.21 (4H, br s), 7.87 (1H, s).

(3)7β-Amino-3-[3-carbamoylamino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylatedihydrochloride

IR (Nujol): 3300, 1770, 1700, 1560 cm⁻¹

NMR (DMSO-d₆, δ): 3.41 (2H, br s), 3.6-3.8 (4H, m), 5.20 (2H, m), 5.43(2H, s), 6.85 (1H, d, J=3 Hz), 8.20 (1H, d, J=3 Hz).

(4)7β-Amino-3-[3-amino-2-(2-carbamoyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylatetrihydrochloride

(5)7β-Amino-3-[2-methyl-3-methylamino-1-pyrazolio]methyl-3-cephem-4-carboxylatetrihydrochloride

NMR (D₂ O--NaHCO₃, δ): 2.93 (3H, s), 3.25-3.38 (2H, m), 3.63 (3H, s),5.07-5.33 (4H, m), 5.97 (1H, d, J=2 Hz), 7.89 (1H, d, J=2 Hz).

(6)7β-Amino-3-[2-methyl-3-methoxycarbonylmethylamino-1-pyrazolio]methyl-3-cephem-4-carboxylatetrihydrochloride

IR (Nujol): 3200-3350, 1780, 1610 cm⁻¹.

EXAMPLE 1

To a solution of7β-amino-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylatetrihydrochloride (2 g) and N-(trimethylsilyl)acetamide (5.85 g) intetrahydrofuran (40 ml)was added(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetyl chloridehydrochloride (1.26 g) under ice-cooling. After being stirred for 1 hourat the same temperature, the reaction mixture was poured intodiisopropyl ether (200 ml). The resulting precipitate was collected byfiltration, dissolved in water (100 ml) and adjusted to pH 2.0 with 5%sodium bicarbonate solution. This solution was subjected to columnchromatography on macroporous non-ionic adsorption resin "Diaion HP-20"(Trademark: prepared by Mitsubishi Chemical Industries). The desiredcompound was eluted with 10% isopropyl alcohol and lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer) (1.07 g).

IR (Nujol): 3150, 1760, 1660 cm⁻¹

NMR (DMSO-d₆, δ): 2.90 and 3.23 (2H, ABq, J=18 Hz), 3.68 (2H, br s),4.38 (2H, m), 4.63 (2H, d, J=5 Hz), 5.02 (1H, d, J=5 Hz), 5.09-5.47 (2H,m), 5.04 and 5.33 (2H, ABq, J=15 Hz), 5.64 (1H, dd, J=5, 8 Hz),5.78-6.16 (1H, m), 5.81 (1H, d, J=3 Hz), 7.26 (2H, br s), 8.08 (1H, d,J=3 Hz), 8.10 (2H, br s), 9.50 (1H, d, J=8 Hz).

EXAMPLE 2

The following compounds were obtained according to a similar manner tothat of Example 1.

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-(2-methyl-3-amino-1-pyrazolio)methyl-3-cephem-4-carboxylate (syn isomer)

IR (Nujol): 3580, 3310, 3200, 1760, 1670, 1615, 1580 cm⁻¹

NMR (DMSO-d₆ -D₂ O, δ): 2.97, 3.23 (2H, ABq, J=18 Hz), 3.68 (3H, s),5.01 (1H, d, J=5 Hz), 4.98, 5.28 (2H, ABq, J=18 Hz), 5.66 (1H, d, J=5Hz), 5.85 (1H, d, J=3 Hz), 8.01 (1H, d, J=3 Hz).

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[2-(2-hydroxyethyl)-3-amino-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 3150-3300, 1760, 1580-1660 cm⁻¹

NMR (DMSO-d₆, δ): 2.92, 3.25 (2H, ABq, J=18 Hz), 3.40-3.77 (3H, m), 3.80(3H, s), 4.14-4.70 (2H, m), 4.90-5.42 (2H, m), 5.63 (1H, dd, J=5 Hz, 8Hz), 5.83 (1H, d, J=3 Hz), 7.17-7.57 (2H, br s), 8.08 (1H, d, J=3 Hz),7.93-8.30 (2H, br s), 9.49 (1H, d, J=8 Hz).

(3)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-4-methyl-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 3250, 1760, 1640, 1600 cm⁻¹

NMR (D₂ O, δ): 1.96 (3H, s), 3.03 and 3.33 (2H, ABq, J=18 Hz), 3.69-3.33(2H, m), 4.17-4.41 (2H, m), 4.78 (2H, d, J=5 Hz), 5.03 (1H, d, J=5 Hz),5.04-5.48 (4H, m), 5.72-6.21 (1H, m), 5.81 (1H, d, J=5 Hz), 7.68 (1H,s).

(4)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-[3-carbamoylamino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 3300, 1770, 1700, 1560 cm⁻¹

NMR (D₂ O, δ): 3.07 (1H, d, J=18 Hz), 3.40 (1H, d, J=18 Hz), 3.84 (4H,m), 4.74 (2H, d, J=5 Hz), 5.20 (1H, d, J=5 Hz), 5.25 (2H, s), 5.1-5.5(2H, m), 5.79 (1H, d, J=5 Hz), 5.9-6.4 (1H, m), 6.74 (1H, d, J=3 Hz),8.04 (1H, d, J=3 Hz).

(5)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[3-amino-2-(2-carbamoyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 3300, 1760, 1710, 1590, 1520 cm⁻¹

NMR (D₂ O, δ): 3.07 (1H, d, J=18 Hz), 3.42 (1H, d, J=18 Hz), 4.06 (3H,s), 4.17-4.60 (4H, m), 4.70-5.43 (3H, m), 5.83 (1H, d, J=5 Hz), 5.94(1H, d, J=3 Hz), 7.87 (1H, d, J=3 Hz).

(6)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-[3-amino-2-(2-carbamoyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 3300, 1770, 1710, 1600 cm⁻¹

NMR (D₂ O, δ): 3.00-3.77 (2H, m), 4.27-4.67 (4H, m), 4.67-5.63 (7H, m),5.96 (1H, d, J=5 Hz), 6.06 (1H, d, J=3 Hz), 7.99 (1H, d, J=3 Hz).

(7)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-[2-methyl-3-methylamino-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 3200-3300, 1770, 1670, 1610, 1520 cm⁻¹

NMR (D₂ O, δ): 2.88 (3H, s), 3.58 (3H, s), 3.03-3.33 (2H, m), 4.50-4.87(2H, m), 4.93-5.48 (3H, m), 5.70-5.97 (2H, m), 7.84 (1H, d, J=3 Hz).

(8)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-(2-methyl-3-methylamino-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 3200-3300, 1760, 1660, 1610, 1520 cm⁻¹

NMR (D₂ O, δ): 2.89 (3H, s), 3.58 (3H, s), 4.04 (3H, s), 3.03-3.30 (2H,m), 4.93-5.25 (2H, m), 5.77-5.97 (2H, m), 7.85 (1H, d, J=3 Hz).

EXAMPLE 3

To a solution of di(trifluoroacetic acid) salt of7β-amino-3-[3-acetamido-2-(2-acetoxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(1 g) and N-(trimethylsilyl)acetamide (2.01 g) in dichloromethane (20ml) was added(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetyl chloridehydrochloride (0.520 g) under stirring and ice-cooling. The mixture wasstirred for one hour at room temperature. The reaction mixture was addedto ether under stirring and ice-cooling. The produced amorphous solidwas dried in vacuo and dissolved in water. The aqueous solution wasadjusted to pH 13 with 1N aqueous sodium hydroxide under stirring at-3°˜0° C. and stirred for 2 hours at the same temperature. The aqueoussolution was adjusted to pH 2 with 1N hydrochloric acid and subjected tocolumn chromatography on Diaion HP-20 and the elution was carried outwith 10% aqueous isopropyl alcohol. The fractions containing the objectcompound were combined, concentrated to remove isopropyl alcohol andlyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-[3-acetamido-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer) (222 mg).

mp: 150° C. (dec.)

IR (Nujol): 3200, 1770, 1660, 1600 cm⁻¹

NMR (D₂ O, δ): 2.22 (3H, s), 3.08 (1H,d, J=18 Hz), 3.42 (1H, d, J=18Hz), 3.7-4.0 (4H, m), 4.74 (2H, d, J=6 Hz), 5.20 (1H, d, J=5 Hz), 5.30(2H, s), 5.1-5.5 (2H, m), 5.81 (1H, d, J=5 Hz), 5.8-6.4 (1H, m), 6.85(1H, d, J=3 Hz), 8.11 (1H, d, J=3 Hz).

EXAMPLE 4

To a solution of7β-amino-3-[2-methyl-3-methoxycarbonylmethylamino-1-pyrazolio]methyl-3-cephem-4-carboxylatetrihydrochloride (0.7 g) and N-(trimethylsilyl)acetamide (1.87 g) intetrahydrofuran (14 ml) was added(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetyl chloridehydrochloride (0.37 g) under ice-cooling and the mixture was stirred atroom temperature for 2 hours. The reaction mixture was poured into ethylacetate (140 ml) and the resultant powder was collected by filtration.The powder was dissolved in ice-water and adjusted to pH 13 with 1Naqueous sodium hydroxide. After stirring for 20 minutes underice-cooling, the solution was adjusted to pH 2 with 3N hydrochloric acidand subjected to column chromatography on "Diaion HP-20" using 5%aqueous isopropyl alcohol as an eluent. Fractions containing the objectcompound were combined, evaporated in vacuo to remove isopropyl alcoholand lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-(3-carboxymethylamino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer) (0.24 g).

IR (Nujol): 3250, 1760, 1670, 1610, 1520 cm⁻¹

NMR (D₂ O, δ): 3.10 and 3.38 (2H, ABq, J=18 Hz), 3.67 (3H, s), 3.83 (2H,s), 4.06 (3H, s), 4.97 and 5.28 (2H, ABq, J=14 Hz), 5.18 (1H, d, J=5Hz), 5.82 (1H, d, J=5 Hz), 5.84 (1H, d, J=3 Hz), 7.85 (1H, d, J=3 Hz).

EXAMPLE 5

The following compound was obtained according to a similar manner tothat of Example 4.

7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-(3-carboxymethylamino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 3300, 1770, 1670, 1610, 1520 cm⁻¹

NMR (D₂ O--NaHCO₃, δ): 3.10 and 3.38 (2H, ABq, J=18 Hz), 3.67 (3H, s),3.83 (2H, s), 4.67-4.93 (2H, m), 4.83-5.53 (2H, m), 5.19 (1H, d, J=5Hz), 5.70-6.23 (3H, m), 7.85 (1H, d, J=3 Hz).

EXAMPLE 6

To a suspension of phosphorus pentachloride (0.89 g) in methylenechloride (18 ml) was added(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-cyclopentyloxyiminoacetic acid(1 g) at -15° C. After being stirred for 30 minutes at -10° C.˜-15° C.,the mixture was evaporated in vacuo to remove methylene chloride. Theresidue was suspended in tetrahydrofuran (16 ml), and then added to asolution of7β-amino-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylatehydrochloride dihydrate (1.6 g) and N-(trimethylsilyl)acetamide (5.1 g)in a mixture of N,N-dimethylformamide (16 ml) and tetrahydrofuran (16ml) at -10° C. After being stirred under ice-cooling for 1 hour, themixture was added dropwise to ethyl acetate. The resulting precipitatewas collected by filtration, dissolved in water and adjusted to pH 4with an aqueous solution of sodium bicarbonate. The aqueous solution wassubjected to column chromatography on Diaion HP-20, and eluted with 10%isopropyl alcohol. The fractions containing the object compound werecollected, and evaporated in vacuo to remove isopropyl alcohol. Theaqueous solution was lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-cyclopentyloxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer) (0.16 g).

IR (Nujol): 3300, 1765, 1660 cm⁻¹

NMR (DMSO-d₆, δ): 1.27-2.02 (8H, m), 2.94 and 3.23 (2H, ABq, J=18 Hz),3.67-3.82 (2H, m), 4.20-4.52 (2H, m), 4.57-4.83 (1H, m), 5.01 (1H, d,J=5 Hz), 5.02 and 5.28 (2H, ABq, J=15 Hz), 5.61 (1H, dd, J=5 Hz, 8 Hz),5.81 (1H, d, J=3 Hz), 7.31 (2H, br s), 8.06 (1H, d, J=3 Hz), 8.07 (2H,br s), 9.38 (1H, d, J=8 Hz).

EXAMPLE 7

7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-cyclobutoxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer) was obtained according to a similar manner to that ofExample 6.

IR (Nujol): 3250, 1765, 1660 cm⁻¹

NMR (DMSO-d₆, δ): 1.45-1.70 (2H, m), 2.10-2.30 (4H, m), 2.99 and 3.20(2H, ABq, J=18 Hz), 4.20-4.60 (4H, m), 4.71 (1H, m), 4.90-5.40 (2H, m),5.04 (1H, d, J=5 Hz), 5.65 (1H, dd, J=8 Hz, 5 Hz), 5.82 (1H, d, J=3 Hz),7.38 (2H, br s), 8.10 (1H, d, J=3 Hz), 8.19 (2H, br s), 9.53 (1H, d, J=8Hz).

EXAMPLE 8

A solution of7β-amino-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylatehydrochloride (50 g) in a mixture of acetonitrile (500 ml) and water(500 ml) was adjusted to pH 6.0 with aqueous sodium bicarbonatesolution. To the resulting solution was added(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetyl chloridehydrochloride (40 g) under stirring at 5° C. and the mixture was stirredat the same temperature for 1.5 hours, keeping pH 5.0-6.5 with aqueoussodium bicarbonate solution. The reaction mixture was evaporated toremove the containing organic solvent. The water solution was subjectedto column chromatography on Sepabeads SP-205 (Trademark: MitsubishiKasei Corporation) (1.3 l). The column was washed with water (5 l) andthe object compound was eluted with 15% isopropyl alcohol, and theeluate was lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer) (51.2 g).

IR (Nujol): 3300, 1750, 1660, 1600 cm⁻¹

NMR (DMSO-d₆, δ): 1.23 (3H, t, J=7 Hz), 2.93 and 3.18 (2H, ABq, J=18Hz), 3.58 (2H, br s), 4.15 (2H, q, J=7 Hz), 4.13-4.25 (2H, m), 5.01 (1H,d, J=5 Hz), 5.03 and 5.23 (2H, ABq, J=16 Hz), 5.63 (1H, dd, J=5 Hz, 8Hz), 5.81 (1H, d, J=3 Hz), 7.27 (2H, br s), 8.10 (1H, d, J=3 Hz), 8.15(2H, br s), 9.50 (1H, d, J=8 Hz).

EXAMPLE 9

The following compounds were obtained according to a similar manner tothat of Example 8.

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(n-propoxyimino)acetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 3350, 3130, 1765, 1610 cm⁻¹

NMR (DMSO-d₆, δ): 0.84-0.93 (3H, m), 1.53-1.73 (2H, m), 3.00 and 3.22(2H, ABq, J=18 Hz), 3.60 (2H, br s), 4.00-4.10 (2H, m), 4.15-4.30 (2H,m), 5.05 (1H, d, J=5 Hz), 5.06 and 5.25 (2H, ABq, J=16 Hz), 5.69 (1H,dd, J=5 Hz, 8 Hz), 5.84 (1H, d, J=3 Hz), 7.40 (2H, br s), 8.08 (1H, d,J=3 Hz), 8.17 (2H, br s), 9.52 (1H, d, J=8 Hz).

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-methyl-2-propenyloxyimino)acetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 1755 cm⁻¹.

(3)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(2-methyl-2-propenyloxyimino)acetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol): 1760 cm⁻¹.

EXAMPLE 10

To a solution of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer) (30 g) in 4N-hydrochloric acid (22.4 ml) was added acetone(120 ml) under stirring at 20° C. The stirring was continued for 10hours at the same temperature to give crystals, which were collected byfiltration and dried to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylatehydrochloride (syn isomer) (13.46 g).

IR (Nujol): 3250, 3150, 3120, 1785, 1705, 1650, 1620, 1575, 1530 cm⁻¹

NMR (DMSO-d₆, δ): 1.23 (3H, t, J=7 Hz), 3.24 and 3.30 (2H, ABq, J=18Hz), 3.60 (2H, br s), 4.03-4.25 (2H, m), 4.13 (2H, q, J=7 Hz), 5.14 and5.32 (2H, d, J=16 Hz), 5.18 (1H, d, J=5 Hz), 5.86 (1H, dd, J=5 Hz, 8Hz), 5.89 (1H, d, J=3 Hz), 7.50 (2H, br s), 8.03 (1H, d, J=3 Hz), 8.21(2H, br s), 8.58 (1H, d, J=8 Hz).

EXAMPLE 11

To a solution of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer) (5 g) in water (50 ml) was added 1M sulfuric acid (8 ml).The solution was lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylatesulfate (syn isomer) (5.2 g).

IR (Nujol): 3150, 1760, 1640, 1610 cm⁻¹

NMR (DMSO-d₆, δ): 1.23 (3H, t, J=7 Hz), 3.21 and 3.32 (2H, ABq, J=18Hz), 3.62 (2H, br s), 4.03-4.26 (2H, m), 4.13 (2H, q, J=7 Hz), 5.88 (1H,dd, J=5 Hz, 8 Hz), 5.90 (1H, d, J=3 Hz), 7.50 (2H, br s), 8.04 (1H, d,J=3 Hz), 8.21.(2H, br s), 8.59 (1H, d, J=8 Hz).

What we claim is: 1.7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate(syn isomer) or its acid addition salt.
 2. A pharmaceutical compositionwhich comprises, as an active ingredient, an effective amount of acompound of claim 1 or a pharmaceutically acceptable salt thereof inadmixture with a pharmaceutically acceptable carrier.
 3. A method forthe treatment of infectious diseases which comprises administering aneffective amount of a compound of claim 1 or a pharmaceuticallyacceptable salt thereof to a human or animal.